雷帕霉素通过诱导自噬抑制子宫内膜癌HEC-1B细胞的恶性生物学行为研究

The study of rapamycin inhibiting the malignant biological behavior of endometrial carcinoma HEC-1B cells by inducing autophagy

目的研究雷帕霉素对人子宫内膜癌HEC-1B细胞增殖、侵袭迁移能力的影响及其可能的机制。方法不同浓度雷帕霉素作用于HEC-1B细胞不同时间后,采用CCK-8法检测细胞增殖能力并计算雷帕霉素的半数抑制浓度(IC50),划痕及Transwell试验检测细胞侵袭迁移能力;在激光共聚焦显微镜下观察LC3颗粒的聚集情况;Western blot检测对照组及雷帕霉素组细胞中LC3Ⅱ、p62蛋白表达情况。结果在一定药物浓度及时间条件下,雷帕霉素抑制HEC-1B细胞的生长活性,且与药物浓度和作用时间呈正相关(P<0.01);雷帕霉素呈浓度依赖性的抑制细胞侵袭及迁移能力(P<0.05);在激光共聚焦显微镜下可见细胞经雷帕霉素处理后出现较明显的LC3颗粒聚集;雷帕霉素能升高LC3Ⅱ蛋白表达量(P<0.05),降低p62蛋白表达量(P<0.05)。结论雷帕霉素可通过诱导HEC-1B细胞自噬抑制子宫内膜癌细胞的增殖、侵袭迁移能力。

Objective To observe the effects of rapamycin on the proliferation and migration of human endometrial cancer HEC-1B cells,and explore its possible mechanism.Methods In the condition of different concentrations of rapamycin on HEC-1B at different times,CCK-8 method was used to detect the cell proliferation ability and calculate the half inhibitory concentration(IC50)of rapamycin,the scratch and Transwell test were used to detect the invasion and migration ability of cells;the aggregation of LC3 particles was observed under a laser confocal microscope,Western blotting method was used to detect the expression of LC3 Ⅱ and p62 in the control group and rapamycin group.Results Under certain drug concentration and time conditions,rapamycin inhibited the growth activity of HEC-1B cell,and was positively correlated with drug concentration and duration of action(P<0.01);the migration distance and the number of transmembrane cells were smaller than those of the control group(P<0.05),and the degree of inhibition gradually increased with the increase of drug concentration;under the laser confocal microscope,it was seen that cells treated with rapamycin showed more obvious LC3 particle aggregation;rapamycin increased the expression of LC3 Ⅱ protein(P<0.05),and decreased the expression of p62 protein(P<0.01).Conclusion Rapamycin can inhibit the proliferation,invasion and migration of endometrial cancer cells by inducing HEC-1B cell autophagy.